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Re: Ask Quick Questions Here [Re: Chamo]
    #28383322 -

I'll make a syringe and put a rubber band around it long ways to create pressure if it's stubborn. Use slightly softer agar. Only had spores like 5 years old done like this, so idk beyond that how well it works

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Re: Ask Quick Questions Here [Re: Smellyhobbit]
    #28383325 -

I’m doing that as well. I’m just taking this opportunity to try various ways to rehydrate them. There’s some information online of spores being hydrated at different pressures.

I just can not find any information on what’s happens when the environment that the spores are in become saturated with the carbon dioxide.

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Re: Ask Quick Questions Here [Re: Chamo]
    #28383328 -

Not sure to be honest. Go for it if you wanna. If the question is what will happen, idk. If the question is “Is there value in the effort?” then probably not tbh. Bit of a reinvent the wheel sort of thing.


--------------------
A Love Letter to New Growers
The 7 Edicts of Mushroom Cultivation as Decreed by Smellyhobbit

:kodama:


Anyone can get a canopy. But can you get what you want?

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Re: Ask Quick Questions Here [Re: Smellyhobbit]
    #28383333 -

I understand you. I’m not expecting anything to happen. I’m doing it the normal way as well.

I was inspired by an article I read online of spores being hydrated at different pressures. The only difference being this pressure would be generated by co2. I wonder if it would just kill the spores

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Re: Ask Quick Questions Here [Re: Chamo] * 1
    #28383341 -

Cool beans. Give it a go and let us know what happens


--------------------
A Love Letter to New Growers
The 7 Edicts of Mushroom Cultivation as Decreed by Smellyhobbit

:kodama:


Anyone can get a canopy. But can you get what you want?

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Re: Ask Quick Questions Here [Re: Smellyhobbit] * 2
    #28383363 -

Seems they’ll be soaking in carbonic acid which may be stabilized in such a small volume of solution under constant pressure of co2.  Im not a chemist so not sure on that but would think it won’t be beneficial to the spores as weak organic acids are used in food preservation to prevent fungal growth.

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Re: Ask Quick Questions Here [Re: Mycolorado]
    #28383370 -

To clarify, I meant that I would keep the spores separate from the reaction. So they would only be exposed to co2 gas, I think.



Something ghetto rigged like this

I didn’t have to buy anything so if it doesn’t work no harm. All of it is probably not going to work with spores that old.

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Re: Ask Quick Questions Here [Re: Chamo] * 1
    #28383372 -

The water is being exposed to co2 which will immediately absorb into the water creating carbonic acid.

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Re: Ask Quick Questions Here [Re: Mycolorado]
    #28383378 -

Ah yes you’re right

It’s been a long time since chemistry class lol

Thanks myco that makes sense

Edited by Chamo (07/03/23 11:25 AM)

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Re: Ask Quick Questions Here [Re: hazyhorse]
    #28383439 -

Will Spores survive  in tap water? Concerned about chlorine. Seems average of .89ppm in my area according to the counties report for 2022 calendar year.

Washed off a bunch of fresh Gymns the other day, kept the water after I saw it turned a rusty orange in hopes of nocc’ing up some logs/stumps.


--------------------
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                    🌕🌖🌗🌘🌑🌒🌓🌔🌕
  :sporedrop:Gymnopilus subspectibilis grow log:sporedrop:             
•89g fresh subspectibilis tea•
•Rapéh Crafters•
• Gift yourself a healthy dose of Self banning•

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From spores to single set of genetics [Re: Nichrome]
    #28383444 -

How many transfers do you find it takes from single drop of spores to single set of genetics if you don’t streak the drop around your agar?

Say you put a drop of a MSS into the center of your agar then transfer the “best/strongest” mycelium from the outermost growth, that mycelium is still germinating from a cluster of millions of spores.

How many transfers does it usually take you to get a uniform set of growth on your agar?
I’m around 4-5 transfers into my first time with agar and half my plates are yet to be very uniform with their growth so I’m assuming I’ve yet to isolate anything very accurately.

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Re: From spores to single set of genetics [Re: MicroMycelium]
    #28383447 -

The only isolating you're doing here is isolation from contamination. I know this forum finds this topic contentious, for some reason, but you're not isolating individual strains or "genetics" by making plate transfers.

Symmetrical growth is an indication of health, it takes as long as it takes. The number of transfers to achieve can vary.

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Re: From spores to single set of genetics [Re: Stipe-n Cap]
    #28383462 -

Stipe-n Cap said:
The only isolating you're doing here is isolation from contamination. I know that this forum finds this topic contentious, for some reason, but you're not isolating individual strains or "genetics" by making plate transfers.

Symmetrical growth is an indication of health, it takes as long as it takes. The number of transfers to achieve can vary.



I was under the impression that MSS allow different combinations of dikaryotic mycelium to form during germination.

A single drop of spores has the potential of forming millions of monokaryotic mycelia which can then mix & match to then later form different fruits.
Physically sectoring off the mycelium should cause separation of those combinations over time…
Unless you are saying every single spore will mate/germination in an orgy of spore-to-mycelium baby making and be forever bonded..?:confused:



Edited by MicroMycelium (07/03/23 12:38 PM)

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Re: From spores to single set of genetics [Re: Stipe-n Cap]
    #28383467 -

Afternoon, stipe.

“Mushroom researchers and cultivators commonly isolate mushroom strains using three methods: tissue isolation, spore isolation, and substrate mycelium isolation. Tissue isolation involves isolating a small portion of the inner tissue of fruit bodies for re-growth to obtain strains of interest, and has been employed for the isolation of the basidiomycetes, G. lucidum [7], Tricholoma matsutake [8], and Pleurotus spp. [9]. Spore isolation is mainly used to obtain single-spore cultures, which can be employed as units for hybridization breeding of mushroom species, including the successful breeding of improved Pleurotus ostreatus and L. edodes strains [10, 11]. Single-spore cultures can also be used for mating system analysis of mushrooms [12, 13]. As for substrate mycelium isolation, strains can be obtained from a substrate in which mushrooms have been grown previously [14]. Of the three methods, tissue isolation is practiced most frequently because of the low contamination rate, high reliability, and ease of manipulation.”

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Re: From spores to single set of genetics [Re: MicroMycelium]
    #28383469 -

What should i do with this gnat infested 3 litre container in which i buried a PF cake 16 days ago?







My heart says keep it, my mind says toss it.

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Re: From spores to single set of genetics [Re: MicroMycelium]
    #28383470 -

So I've got some good rhyzo growth coming in. Looks better in person than in the pics.

Beige plate came from what I thought was the second best plate, but it has the best uniform growth.



These two are others that came from what I thought was the best plate, still way less uniform. I guess I'll still transform them out by grabbing from the thicker rhyzo areas. Or should I not even bother (pretty sure I still will)

This one would be 6 o clock



This one would be 10ish



The only thing I'm wondering is if it's the type of plate. Pink ones are classic pasty's just with a bit less agar, and the beige one was oats boil water with instant coffee. I actually might prefer this recipe.

Also, tips for decreasing condensation inside?

Edited by Huskies (07/03/23 12:40 PM)

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Re: From spores to single set of genetics [Re: Chamo]
    #28383473 -

Chamo said:
Afternoon, stipe.

“Mushroom researchers and cultivators commonly isolate mushroom strains using three methods: tissue isolation, spore isolation, and substrate mycelium isolation. Tissue isolation involves isolating a small portion of the inner tissue of fruit bodies for re-growth to obtain strains of interest, and has been employed for the isolation of the basidiomycetes, G. lucidum [7], Tricholoma matsutake [8], and Pleurotus spp. [9]. Spore isolation is mainly used to obtain single-spore cultures, which can be employed as units for hybridization breeding of mushroom species, including the successful breeding of improved Pleurotus ostreatus and L. edodes strains [10, 11]. Single-spore cultures can also be used for mating system analysis of mushrooms [12, 13]. As for substrate mycelium isolation, strains can be obtained from a substrate in which mushrooms have been grown previously [14]. Of the three methods, tissue isolation is practiced most frequently because of the low contamination rate, high reliability, and ease of manipulation.”



:takingnotes:

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Re: From spores to single set of genetics [Re: MicroMycelium] * 1
    #28383475 -

Yeah, multispore forms many connections, this is the point. Monokaryotic stage is extremely short lived due to high mating compatibility.

Hyphae are microscopic:



Hyphae fuse and branch laterally during apical elongation. Bullers phenomenon connects dikaryons to compatible monokaryons, then grow in close proximity measured in nanometers.

Each "sector" is a woven mat of mycelium containing something like 104(for arguments sake) hyphae of mixed genetic material. Teasing those apart with a scalpel, needle biopsy, etc, is impossible.

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Re: From spores to single set of genetics [Re: Yahra] * 2
    #28383476 -

Yahra said:
What should i do with this gnat infested 3 litre container in which i buried a PF cake 16 days ago?







My heart says keep it, my mind says toss it.



I keep all of them until they contaminate. I say keep it.

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Re: From spores to single set of genetics [Re: Stipe-n Cap]
    #28383482 -

After a lot of reading I concede that it would be very hard, if not impossible without new instruments to practically achieve a genetic isolate with a scalpel. But it can be done in labs. I was incorrect on my assumption of how easy it is. But I would still prefer a culture as isolated as much as possible. Don’t you agree that it would give more consistency even if it were not a “true” isolate?

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Shop: Original Sensible Seeds Bulk Cannabis Seeds   Mushroom-Hut Liquid Cultures   Sporeworks.EU Spores for European Microscopy   MagicBag.co Certified Organic All-In-One Grow Bags   Myyco.com Isolated Cubensis Liquid Culture For Sale   North Spore Bulk Substrate


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